Abcam's Glucose 6 Phosphate Dehydrogenase Assay Kit (Colorimetric) (ab102529) is a simple, sensitive and rapid assay detects the activity of Glucose 6 Phosphate Dehydrogenase (G6PDH) in a variety of samples. In the assay, glucose 6 phosphate is oxidized, which leads to the conversion of a nearly colorless probe to an intensely colored product with an absorbance at 450nm.

RelevanceGlucose 6 phosphate dehydrogenase (G6PDH) is a cytosolic enzyme in the pentose phosphate pathway, a metabolic pathway that supplies reducing energy to cells (such as erythrocytes) by maintaining the level of the co-enzyme nicotinamide adenine dinucleotide phosphate (NADPH). The NADPH in turn maintains the level of glutathione in these cells that helps protect the red blood cells against oxidative damage. Of greater quantitative importance is the production of NADPH for tissues actively engaged in biosynthesis of fatty acids and/or isoprenoids, such as the liver, mammary glands, adipose tissue, and the adrenal glands.

Methanol lysis holds advantages over both NP-40 and mechanical lysis in that it much quicker, requiring only 2 minutes incubation at room temperature, followed by centrifugation and aspiration of the supernatant. Please find attached data from my experiment which shows that, despite identical NADH standard curves, lysis of human PBMCs in methanol leads to loss of measurable G6PDH activity when compared to NP-40 lysis.

We have not tested this specific combination of components in the concentrations mentioned. So there can always be some unforeseen interference, but theoretically the composition should work with this kit.

We have not tried using blood samples with this assay kit. Since blood is coloured, it will interfere with the colour of the reaction and contribute to the final readings. Also, since this is an enzymatic assay, you cannot use a filter to remove the pr...

For cells with this assay, 2-5 X 10^6 cells can be homogenized with an equivalent volume of ice cold PBS or other buffer (pH 6.5 - 8). Add 1 - 50µl samples into duplicate wells of a 96-well plate and bring volume to 50 µl with Assay Buffer....